Method for obtaining male-sterile plants

Multicellular living organisms and unmodified parts thereof and – Plant – seedling – plant seed – or plant part – per se – Higher plant – seedling – plant seed – or plant part

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800274, 800278, 800286, 800287, 800298, 435419, 435468, 536 236, 536 241, 536 245, A01H 500, A01H 102, C12N 1529, C12N 1582

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059327847

DESCRIPTION:

BRIEF SUMMARY
FIELD OF THE INVENTION

The invention involves genetic manipulation of plants by the application of recombinant DNA technology. The invention describes a method for obtaining plants which display nuclear encoded male-sterility, due to expression of said recombinant DNA as well as parts of said plants which are either sexually or asexually reproducible, or both.


BACKGROUND OF THE ART

Heterosis is the effect that progeny obtained through cross-pollination show better agronomical properties than progeny derived through self-pollination. These better properties result in a higher profit which is the reason that the development of hybrid seed is one of the prime objectives for the seed industry.
To obtain hybrid seeds it is necessary that the female plants are unable to self-pollinate. Many crop plants accommodate both the female and male reproductive organs on the same individual and thus self-pollination is dominating cross-pollination. Introduction of male sterility in many crops is thus necessary for hybrid seed production.
In production fields the male-sterile acceptor and male-fertile donor plants are grown side by side and after cross-pollination the hybrid seed is harvested. Hybrid seed is separately collected from the non-hybrid seeds formed on the donor plants by destruction of these male-fertile plants before the harvest. This approach makes the discrimination between fertile and sterile plants necessary which can be accomplished by the appearance of different phenotypes. Also a selectable marker gene such as a gene coding for a herbicide resistance, closely linked to the male-sterility locus can be used for this purpose. Selection of the hybrid seeds can than be accomplished by spraying the herbicide, which will result in the destruction of the male-fertile plants. Alternatively the hybrid seeds can be selected after harvesting by a phenotypic marker expressed at the seed level.
Nowadays in agricultural practice male-sterile parental lines are obtained by physically emasculation of the plants or by the application of cytoplasmic or nuclear encoded male-sterile mutants. There are only a limited number of natural male-sterile mutants available in the commercial interesting crops. This renders the latter approach often not feasible. In addition naturally male-sterile plants have their disadvantages. There preparation is laborious and there maintenance and propagation difficult. They often show additional, detrimental traits and a difficult inheritance.
The development of nuclear encoded male-sterile plants by the application of recombinant DNA techniques has succeeded in new approaches which circumvent most of the disadvantages mentioned above.


STATE OF THE ART

The International Patent Application WO92/18625, MOGEN International N.V. proposes methods for the production of restorable male-sterile plants in general terms, essentially comprising integration of a recombinant polynucleotide into their genome, essentially comprising an inhibitory gene, which, upon proper expression in the anthers of the plant, is capable of inhibiting expression of one or more genes encoding an enzyme involved in the synthesis of chalcone, or one of its precursors.


DEFINITIONS

Antisense gene: a nucleotide sequence having a homology of more than 50%, preferably more than 80% with a target gene as defined herein, and which is linked to a promotor in 3' to 5' orientation with respect to the target gene and can be expressed as an RNA molecule.
Gene or sense gene: a nucleotide sequence that can be expressed as RNA molecule and/or polypeptide.
Promoter: a nucleotide sequence which directs the expression of a (sense-) gene or antisense gene, or nucleotide sequences derived thereof.
Inhibitor gene: a (sense-)gene or antisense gene, expression of which leads to prevention or inhibition of the expression of a target gene as defined herein.
Repair gene: a gene capable to prevent or sufficiently inhibit the action of an inhibitor gene thus repairing the activity of a target gene.
Target gene: a gene which activity is to be inhi

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